Cell culture and tissue engineering are critical technologies in regenerative medicine, which provides sufficient cells for experimental analysis through large-scale proliferation by artificial means, and then conditions required for the growth and development of tissue cells are simulated and provided, thereby enabling cells obtained by culture to grow and differentiate into cells or tissues with specific characteristics.
Regenerative medicine provides solutions to problems such as uncertainties of organ donation and potential graft rejections due to immune responses following organ transplantation. However, its development is limited by a few techniques, for example, cell culture and three dimensional scaffolds.
In the field of biological cells, it is generally believed that the biomimetic activity of a three-dimensional culture is better than that of a two-dimensional monolayer culture. A number of three-dimensional cell culture methods have thus developed, such as a hydrogel, a suspension, a hanging drop culture, a micromass culture, and a non-adherent substrate. In the field of cell culture, in order for the cultured cells to grow into tissues or organs with desired functions and forms, the use of scaffolds plays an important role. The function of the scaffold is to provide a three-dimensional framework suitable for cell growth, which is commonly known as a three-dimensional scaffold. It has a large number of pores for cell attachments or inoculations, guiding the cells to grow and differentiate in three-dimensional directions as planned to produce simulated and regenerated tissues or organs.
In traditional flat cell culture, there is only a very small area of contact between cells, half of the surface area of a cell is in contact with the culture plate, and the other half is in contact with the culture medium. A three-dimensional culture environment provides other advantages, it is capable of: providing better biochemical signals to direct cell functions, allowing cell migration within the scaffold, increasing cell density and increasing signal transmission among cells, providing molecules for cell attachments and for inducing cell differentiation. When the pore size of a sponge-like three-dimensional scaffold is greater than 50 μm, cell migration is enhanced and more uniform distribution of seeded cells and nutrients are facilitated by the inter-connecting porous structure.
Therefore, one of the important issues in the field is to provide a method and a device for preparing a three-dimensional scaffold, which can achieve the purpose of preparing the three-dimensional scaffold in a simple manner with simple devices, allowing cells to be cultivated stably and grow eventually into tissues or organs with desired functions and forms.
In view of the foregoing, an object of the present invention is to provide a method for preparing a cellulose sponge and a mixed solution thereof, which can achieve the purpose of preparing the three-dimensional scaffold in a simple manner and in a solution, thereby simplifying the complicated process for preparing the scaffold and shortening the required preparation time.